KR20180008352A - Method for producing ginseng extract with enhanced ginsenoside Rg3, F1, C-K, Rh2 and PPT content - Google Patents

Abstract

The present invention relates to a method of preparing a ginseng extract having an increased content of ginsenoside Rg3, F1, C-K, Rh21 and PPT, wherein the method comprises the steps of: enzymatically treating ginseng and drying the same; puffing the dried ginseng; and extracting the puffed ginseng. In the ginseng extract prepared according to the present invention, ginsenoside Rb1, Rb2, Rc, Rd and Re, which are ginsenosides of low bioavailability contained in ginseng, are converted into ginsenosides of high bioavailability, which are Rg3, F1, C-K, Rh21 and PPT, thus providing excellent antioxidant and wrinkle-reducing effects on the skin.

Description

(Method for producing ginseng extract with enhanced ginsenoside Rg3, F1, C-K, Rh2 and PPT content) in which the content of ginsenosides Rg3, F1, C-

The present invention relates to a method for producing ginseng extract having enhanced contents of ginsenosides Rg3, F1, CK, Rh21 and PPT.

Ginseng ( Panax ginseng CA Meyer ) is a herb that belongs to the genus Ogawa and Ginseng. It has been used in Korea, China and Japan for over 2,000 years and has been used to prevent disease and prolong its life. The effects of ginseng on the central nervous system, anticancer activity, anti-cancer activity, immune function regulation, anti-diabetic effect, hepatic function hyperactivity, cardiovascular disorder improvement, anti-arteriosclerosis effect, blood pressure control effect, , Osteoporosis improvement, anti-stress action, anti-fatigue action, antioxidant activity, and inhibition of aging.

Ginseng contains more than 40 kinds of ginsenosides (glycosides) as physiologically active substances of non saponin type, including polyacetylene, phenolic compounds, acid polysaccharides, peptides Peptides, and Alkaloids.

Ginsenoside, a representative physiologically active ingredient of ginseng, refers to saponin contained in ginseng, and ginseng contains various kinds of ginsenosides. Ginsenosides are distributed evenly in the ground and underground of ginseng. Especially, it is known that not only ginsenoside content but also composition is different depending on the parts such as ginseng roots, ginseng leaves, and ginseng fruit.

The ginsenosides of ginseng constitute about 5% of the ginseng dry matter. The Ginsenosides of the ginseng contain about 5% of the ginseng in the Triterpenoid series of Dammarane skeleton containing glucose (Glucose), Arabinose, Xylose, (Rhamnose) and the like. (3, 6, 12 positions) Protopanaxadiol (PPD) saponins, 3 individuals (positions 3, 6 and 12) in the case of two individuals (positions 3 and 12) (Protopanaxatriol; PPT) family saponins. Glucose, Arabinose, Xylose, Rhamnose and the like are bonded to -OH in the R1, R2 and R3 positions of PPD and PPT. More than 30 kinds of ginsenosides have been reported to be isolated from ginseng. However, 6 kinds of Rb1, Rb2, Rc, Rd, Re and Rg1 account for more than 90% The remaining saponin content is low.

The ginsenosides are not absorbed as they are in the human body, but they are not absorbed in the body but are present in the human body, such as Bacteriodes , Lactobacillus , Fusobacterium , Eubacterium , Provetella , It is degraded by microorganisms such as species and the metabolite is absorbed. In the case of ginsenoside Rb1, it is sequentially converted to ginsenoside Rd, compound K (hereinafter abbreviated as CK) and protopanaxadiol by intestinal microorganisms. Rb1, Rb2, Rc, and Rd, which are PPD-derived ginsenosides, are metabolized and absorbed by 20-O- β-D-glucopyranosyl-20 (S) -protopanaxadiol (CK) in the intestine. PPT ginsenosides Re, Rf, and Rg1 are metabolized in the intestine to Rh1, F1, and Protopanaxatriol. The PPD family ginsenosides Rb1, Rb2, Rc and Rd are converted into 20 (R) - / 20 (S) -ginsenoside Rg3 by intestinal microorganisms and Rg3 is converted into Rh2 by intestinal microorganisms.

In recent years, the effective ingredient of ginseng is easily absorbed into the human body, and a method of fermenting ginseng using intestinal microorganism or lactic acid bacteria or treating an enzyme with a trace amount of ingredients in ginseng is used. Korean Patent Publication No. 10-1017378 discloses a method for producing fermented red ginseng and fermented red ginseng and a Bacillus subtilis strain derived from chungkukjang, which is suitable for the fermented ginseng and fermented red ginseng, using a microorganism belonging to the genus Bacillus isolated from chungkukjang. And fermenting red ginseng to increase the content of ginsenosides Rd. The present invention also relates to a method for producing fermented ginseng and fermented red ginseng, and a technology for producing Bacillus subtilis strain derived from chungkukjang.

Korean Patent Registration No. 10-1240192 discloses a fermented ginseng extract for improving insulin secretion using Lactobacillus acidophilus strain K-59 and strain K-59 and a method for producing the same. Rg1, Rb1, Rg1 or Re, which is a low bioavailability component of ginseng, is converted into ginsenoside Rh1, Rg3, Rh2 or CK ( Lactobacillus acidophilus ) by using Lactobacillus acidophilus strain K-59 (KFCC11467P) A method for converting the above-mentioned compound is disclosed.

Accordingly, the present inventors have found that when an extract is prepared by optimally mixing a specific region of ginseng and subjecting it to enzymatic and pelletizing treatment, ginsenosides Rb1, Rb2, Rc, Rd and Re can be converted into ginsenosides Rg3, F1, CK, Rh21 and PPT having high bioavailability, thus giving skin antioxidant and wrinkle-improving effect, and the present invention has been completed.

Korean Patent Publication No. 10-1017378 Korean Patent Publication No. 10-1240192

It is an object of the present invention to provide a method for producing ginseng extract having enhanced contents of ginsenosides Rg3, F1, C-K, Rh21 and PPT.

Another object of the present invention is to provide a ginseng extract having enhanced contents of ginsenosides Rg3, F1, C-K, Rh21 and PPT.

It is another object of the present invention to provide a cosmetic composition comprising ginseng extract.

In order to accomplish the above object, the present invention provides a method for producing ginseng, comprising: enzymatically treating and drying ginseng; Expanding dried ginseng; And extracting the expanded ginseng, wherein the content of ginsenosides Rg3, F1, C-K, Rh21 and PPT is increased.

The ginseng according to the present invention is characterized in that leaf, fruit and root are mixed at a mixing ratio of 0.5-1.0: 0.5-1.0: 0.5-5.0, respectively.

In the present invention, the enzyme is characterized by being a pectinase and a cellulase.

In the present invention, lactose is further added during the enzyme treatment.

In the present invention, the enzyme treatment is performed by adding 0.1 to 1.0 part by weight of pectinase and 0.5 to 1.0 part by weight of a cellulase to 100 parts by weight of ginseng, And the reaction is carried out.

In the present invention, the expanding step is performed at a pressure of 5.0 to 10.0 kgf / cm < ² >.

In the present invention, the extracting step is performed by extracting expanded ginseng with ethanol (EtOH) to prepare an extract, followed by fractionation using HP20 porous resin and ethanol (EtOH).

The present invention also provides a ginseng extract prepared through the above-described method.

In the present invention, the ginseng extract has a skin antioxidant and wrinkle-reducing effect.

The present invention also provides a cosmetic composition comprising the ginseng extract.

The ginseng extract prepared from the production method of the present invention is characterized in that ginsenosides Rb1, Rb2, Rc, Rd and Re which are contained in ginseng have low bioavailability, ginsenosides Rg3, F1, CK, Rh21, and PPT, and is excellent in skin antioxidant and wrinkle-reducing effects.

FIG. 1 is a diagram showing the bioconversion process of ginsenoside in the treatment of ginseng with enzyme.
FIG. 2 is a diagram showing a biosynthesis process of ginsenoside in the case of enzyme and expansion treatment of ginseng. FIG.
3 is a graph showing the DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging activity of the ginseng extract obtained from Example 1 and Comparative Example 1 of the present invention.
4 is a graph showing the MMP-1 (Matrix Metallopeptidase-1) inhibitory activity of the ginseng extract obtained from Example 1 and Comparative Example 1. Fig.

Accordingly, the present inventors have found that when ginseng extract is prepared by optimally mixing specific parts of ginseng and treating it with an enzyme and expanding it, ginsenosides Rb1, Rb2, Rc, and Rd which are low in bioavailability contained in ginseng And Re could be converted to bioavailable ginsenosides Rg3, F1, CK, Rh21 and PPT.

In this example, the leaves, fruits and roots of ginseng were controlled at various mixing ratios, and the enzymes and bulging treatment were sequentially carried out, and then the ginseng extract was prepared to confirm the bioconversion ability of the specific ginsenosides. As a result, ginsenosides Rb1, Rb2, Rc, Rd and Re, which are low in bioavailability contained in ginseng, are converted to bioavailable ginsenosides Rg3, F1, CK, Rh21 and PPT And the present invention has been completed.

Accordingly, in one aspect, the present invention provides a method for producing ginseng, comprising: enzymatically treating and drying ginseng; Expanding dried ginseng; And extracting the expanded ginseng. The present invention also relates to a method for producing ginseng extract having enhanced contents of ginsenosides Rg3, F1, C-K, Rh21 and PPT.

A more detailed method of preparing ginseng extract is as follows.

In the present invention, the term "ginseng extract" means a product obtained by sequentially performing the enzymes and bulging treatment of ginseng and extracting it.

First, the variety of the ginseng used in the production of ginseng extract is not particularly limited, but it is preferable to use at least one selected from the group consisting of ginseng, ginseng,

When using the ginseng, it is preferable that the leaves, fruits, and roots of the ginseng, not the ginseng outpast (leaves, roots, and stem) are used at one time, and that the ginsengs are mixed individually at a specific ratio. Rinsing ginsenosides Rb1, Rb2, Rc, Rd, and Re with low bioavailability, when blended at a mixing ratio of 0.5-1.0: 0.5-1.0: 0.5-5.0, It is preferable from the viewpoint that it can be converted into high-bioavailability ginsenosides Rg3, F1, CK, Rh21 and PPT. If the blending ratio of the leaves, fruits and roots of the ginseng is out of the above range, the saponin series is not suitable in composition ratio, and the skin antioxidant and wrinkle improving effect are not sufficiently exhibited.

According to a preferred embodiment of the present invention, when the ginseng is enzymatically treated, the use of pectinase and cellulase increases the content of Ginsenoside Rg3, F1, CK, Rh21 and PPT . Further, it is more preferable to add lactose in order to further increase the conversion ability to ginsenosides having high bioavailability upon enzyme treatment.

The enzymatic treatment is preferably performed by adding 0.1-1.0 parts by weight of pectinase and 0.5-1.0 parts by weight of a cellulase to 100 parts by weight of ginseng and then reacting at 37 to 50 ° C for 3 to 7 days . If 0.1 part by weight and less than 0.5 part by weight of pectinase and cellulase are added to 100 parts by weight of ginseng, respectively, the content of enzyme is relatively low and thus the effect of the enzyme is insignificant And when the addition amounts of pectinase and cellulase are 1.0 parts by weight and 1.0 part by weight, respectively, it is uneconomical because it can exhibit sufficient efficacy without adding more than 1.0 part by weight.

In addition, when the enzyme is treated, the reaction at 37 to 50 ° C for 3 to 7 days is preferable in that the content of ginsenoside Rd, Rg3 and Rg2 can be maximized. If the temperature and time are out of the above range, most of the saponin is undesirably monodispersed.

When the ginseng is enzymatically treated under the above conditions, the sugar of the ginsenosides Rb1, Rb2 and Rc contained in the ginseng extract is hydrolyzed to be converted into ginsenoside Rd, which is again hydrolyzed (Ginsenoside) Rg3, and in the case of Ginsenoside Re, it is further converted to Rg2 (Figure 1).

As described above, the ginseng is subjected to enzymatic treatment and then dried. However, the method is not particularly limited as long as it can dry the enzyme-treated ginseng. However, if the ginseng is dried by hot air drying at 55 to 65 ° C for 48 to 72 hours, It is preferable from the economical point of view.

Next, the dried ginseng is expanded. It is preferable to perform the operation at a pressure of 5.0 to 10.0 kgf / cm ² in terms of expanding the pores and increasing the surface area. If puffing of ginseng is performed at a pressure of less than 5.0 kgf / cm ² , the surface of the tissue can not be increased due to insufficient expansion of the tissue. If the pressure exceeds 10.0 kgf / cm ² It is not preferable since carbonized and harmful components may be produced.

When the ginseng is expanded after enzymatic treatment under the above conditions, Ginsenoside Rd is converted to F2, which is once again converted to C-K. Also, in the case of ginsenoside Re, it is converted to Rg1 and Rg2, and Rg1 is converted to F1 and (R, S) -Rh1, which is once again converted to PPT. Also, Ginsenoside Rg2 is converted to (R, S) -Rh1, which is once again converted to PPT. In the case of ginsenoside Rh21, it is specifically produced when these two processes are combined. That is, a large amount of Ginsenoside Rb1, Rb2, Rc, Rd, and Re is converted into Rg2, Rg3, and CK by enzymatic conversion and then subjected to bulge treatment to obtain Ginsenoside Rg3, F1, CK, PPT (Figure 2).

The expanded ginseng is extracted with ethanol (EtOH) to prepare an extract, and fractionated using HP20 porous resin and ethanol (EtOH) to obtain a final ginseng extract.

As described above, when an extract is prepared by optimally mixing specific parts of ginseng and subjecting it to enzymatic treatment and expansion treatment, ginsenosides Rb1, Rb2, Rc, Rd, Re can be converted into ginsenosides Rg3, F1, CK, Rh21 and PPT having high bioavailability and can give skin antioxidant and wrinkle-reducing effect. Therefore, the ginseng extract was applied to the cosmetic base as an effective ingredient It can be applied as a cosmetic composition.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the scope of the present invention. It will be self-evident.

≪ Examples 1-3 > Preparation of ginseng extract

Example 1

Among the varieties of ginseng ( Panax ginseng CA Meyer ), 100 g of leaf, fruit and root were mixed at a mixing ratio of 0.5: 1: 2 to prepare 100 g of pectinase and 0.1 g of cellulase, respectively. And 0.5 g. The mixture was allowed to react at 50 캜 for 72 hours, and then dried with hot air at 60 캜 for 72 hours. 100 g of the dried ginseng was expanded by heat treatment at 150 kg / cm ² and 150 ° C for 30 minutes using a high-temperature high-pressure treatment apparatus (Jisico, Korea). After expanding, the extracted ginseng was extracted with 70% ethanol (EtOH) at 80 ° C for 3 hours to prepare an extract. The extracted extract was loaded on a column packed with HP20 and purified by sequentially adding ethanol (EtOH) to each concentration (40%, 50%, 65%). Then, the solution eluted at 65% The extracts were concentrated and powdered to prepare final ginseng extract.

Example 2

The leaves, fruits and roots of Panax ginseng CA Meyer were mixed at a mixing ratio of 0.5: 1: 2, respectively, to prepare 100 g of pectinase, cellulase, Lactose ) Were added, and reacted at 50 ° C for 72 hours, followed by hot air drying at 60 ° C for 72 hours. 100 g of the dried ginseng was expanded by heat treatment at 150 kg / cm ² and 150 ° C for 30 minutes using a high-temperature high-pressure treatment apparatus (Jisico, Korea). After expanding, the extracted ginseng was extracted with 70% ethanol (EtOH) at 80 ° C for 3 hours to prepare an extract. The extracted extract was loaded on a column packed with HP20 and purified by sequentially adding ethanol (EtOH) to each concentration (40%, 50%, 65%). Then, the solution eluted at 65% The extracts were concentrated and powdered to prepare final ginseng extract.

Example 3

Among the varieties of ginseng ( Panax ginseng CA Meyer ), 100 g of leaf, fruit and root were mixed at a mixing ratio of 0.5: 0.5: 0.5 to prepare 0.1 g of pectinase and 0.1 g of Cellulase, respectively. And 0.5 g. The mixture was allowed to react at 50 캜 for 72 hours, and then dried with hot air at 60 캜 for 72 hours. 100 g of the dried ginseng was expanded by heat treatment at 150 kg / cm ² and 150 ° C for 30 minutes using a high-temperature high-pressure treatment apparatus (Jisico, Korea). After expanding, the extracted ginseng was extracted with 70% ethanol (EtOH) at 80 ° C for 3 hours to prepare an extract. The extracted extract was loaded on a column packed with HP20 and purified by sequentially adding ethanol (EtOH) to each concentration (40%, 50%, 65%). Then, the solution eluted at 65% The extracts were concentrated and powdered to prepare final ginseng extract.

<Comparative Examples 1 and 2> Preparation of ginseng extract

Comparative Example 1

Among the varieties of Panax ginseng CA Meyer , 100 g of Ginseng ginseng was heat treated at a pressure of 10 kgf / cm ² at 150 ° C for 30 minutes using a high temperature and high pressure treatment device (Jisico, Korea) Ethanol (EtOH) at 80 ℃ for three hours to obtain ginseng extract.

Comparative Example 2

Among the varieties of Panax ginseng CA Meyer , 0.1% by weight and 0.5% by weight of pectinase and cellulase were added to 100 g of ginseng ginseng and reacted at 50 캜 for 72 hours, followed by incubation at 60 캜 for 72 hours Lt; / RTI &gt; The dried ginseng was extracted and concentrated by ethanol (EtOH) at 80 ℃ for 3 hours to produce ginseng extract.

Test Example 1 High Performance Liquid Chromatography (HPLC)

5 g of water-saturated butanol was added to 1 g of the ginseng extract prepared in Examples 1 to 3 and Comparative Examples 1 and 2, followed by mixing. The supernatant of the butanol mixture was taken and concentrated under reduced pressure. The resulting concentrate was dissolved in methanol (HPLC grade) and filtered through a 0.2 μm membrane filter, and used as a sample for HPLC analysis. HPLC analysis was carried out on a C ₁₈ column (3.0 x 50 mm, ID 5 μm) at UV 203 nm. The mobile phase was graded using Acetonitrile (solvent A) and water (solvent B) and separated at a flow rate of 1.6 ml / min. Gradient conditions of the mobile phase are shown in Table 1.

Time (min) Mobile phase Acetonitrile (solvent A) Water (solvent B) 0.00 17 83 6.00 17 83 9.00 23 77 16.50 23.5 76.5 19.00 31 69 29.00 45 55 31.00 47 53 33.00 90 10 34.00 90 10 35.00 17 83 37.00 17 83

Table 2 shows the results of the changes in the ginsenoside components of Examples 1 to 3 and Comparative Examples 1 and 2.

Example 1 (unit: mg / g) Example 2 (unit: mg / g) Example 3 (Unit: mg / g) Comparative Example 1 (unit: mg / g) Comparative Example 2 (unit: mg / g) Rb1 1.43 0.58 0.99 5.4 1.51 Rb2 0.96 0.85 0.98 2.28 0.89 Rc 1.24 1.09 1.25 2.11 0.77 Rd 5.87 5.96 3.89 10.55 1.54 Re 15.54 14.99 5.15 2.14 3.34 Rf 0.18 0.19 0.12 0.11 0.81 Rg2 8.81 9.14 7.99 2.29 0.51 Rg3 3.34 2.95 5.56 1.49 1.21 Rh1 4.48 5.61 0.98 1.36 0.30 Rh2 0.99 1.41 0.32 0.02 0.85 F1 2.95 3.34 2.21 0.51 0.45 F2 0.98 0.85 0.92 1.5 0.84 C-K 1.15 1.54 1.05 0.01 1.35 PPT 1.41 1.09 0.54 0.22 0.15 Rh21 0.29 0.33 0.15 0 0

As a result, as shown in Table 2, the ginsenoside components of Examples 1 to 3 and Comparative Examples 1 and 2 were analyzed. As a result, The content of ginsenosides Rg3, F1, CK, Rh21 and PPT was found to be significantly higher than that of ginseng extract as a whole.

It was confirmed that ginseng was bioconverted to Ginsenoside Rg3, F1, CK, Rh21 and PPT by enzymes and expansion treatments of Ginsenoside Rb1, Rd and Re.

<Test Example 2> Polysaccharide content analysis

Analysis of the polysaccharide content of the ginseng extracts obtained from Examples 1 to 3 and Comparative Examples 1 and 2 was carried out. Carbazole sulfuric acid method was used for the determination of pectin. Carbazole was used as a control. 5 ml of each of Examples 1 to 3 and Comparative Examples 1 and 2 were quantified in a 250 ml flask and 5 ml of distilled water corresponding to 10 times was added. 2 ml of the supernatant was taken by centrifugation (4 ° C, 9000 rpm, 15 minutes) in a reflux extractor at 80 ° C for 6 hours, and 8 ml of 95% methanol was added thereto. This was centrifuged again at 9000 rpm for 15 minutes. After removing 200 μl of precipitate, 4.8 ml of distilled water was added to dissolve, and 0.5 ml of sulfuric acid and 0.25 ml of carbazole were reacted in 85 water bath for 5 minutes. After 5 minutes, the absorbance was measured at 525 nm using a Multi-reader (Biotec, synergy).

The results of the changes of the polysaccharide contents of Examples 1 to 3 and Comparative Examples 1 and 2 are shown in Table 3.

Example 1 (unit: mg / g) Example 2 (unit: mg / g) Example 3 (Unit: mg / g) Comparative Example 1 (unit: mg / g) Comparative Example 2 (unit: mg / g) Polysaccharide 9.97 11.54 7.84 4.45 2.58

As a result, as shown in Table 3, it was confirmed that Polysaccaride content was increased in Examples 1 to 3 as compared to Comparative Examples 1 and 2.

&Lt; Test Example 3 > Analysis of total phenol content

The total phenol contents of the ginseng extracts obtained from Examples 1 to 3 and Comparative Examples 1 and 2 were analyzed. 2 ml of sodium carbonate was added to 100 ul of each of the ginseng extracts obtained in Examples 1 to 3 and Comparative Examples 1 and 2, followed by addition of 2N Folin and Ciocalteu phenol reagent (Sigma), followed by reaction at room temperature for 30 minutes. After incubation at room temperature for 30 minutes, absorbance was measured at 750 nm. At this time, (+) - catechin hydrate ((+) - catechin hydrate; Sigma) was used as the indicator material.

The results of the changes in total phenol contents of Examples 1 to 3 and Comparative Examples 1 and 2 are shown in Table 4 below.

Example 1 (unit: mg / g) Example 2 (unit: mg / g) Example 3 (Unit: mg / g) Comparative Example 1 (unit: mg / g) Comparative Example 2 (unit: mg / g) Total phenol 27.12 33.51 19.54 4.66 2.81

As a result, as shown in Table 4, it was confirmed that the content of the phenol compound was increased in Examples 1 to 3 as compared to Comparative Examples 1 and 2.

&Lt; Test Example 4 > Cytotoxicity test

The cytotoxicity of the ginseng extracts obtained from Examples 1 to 3 and Comparative Examples 1 and 2 was measured. First, human keratinocyte HaCat (ACTT, CLS 300493, USA) was counted in the same manner as that of a 96-well plate at a density of 2.0 × 10 ⁴ cells / well using a Hemacytometer, Of carbon dioxide for 24 hours. The cell culture obtained after 24 hours of culture was removed, and the ginseng extracts obtained in Examples 1 to 3 and Comparative Examples 1 and 2 were prepared at concentrations of 0, 10, 25, 50, 100 and 200 ug / ml. 1 μl of each concentration was mixed with the DMEM medium, and then treated for each concentration in each well. The cells were cultured at 37 ° C under 5% carbon dioxide for 24 hours. Then, a solution of MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] per well was added at a concentration of 5 mg / ml and further cultured for 4 hours under the same conditions. After 4 hours, the MTT solution was removed and 200 μl of DMSO (dimethyl sulfoxide) solution was added to dissolve the blue formazan. Then, absorbance was measured at 570 nm using a Multi-reader (Biotec, synergy). As a control group, the absorbance was measured using distilled water instead of the sample. The cell survival rate is shown in the following equation (1).

[Equation 1]

Cell survival rate (%) = (absorbance of experimental group / absorbance of control group) X 100

Cell viability (%) density( ug / ml) 0 10 25 50 100 sample Comparative Example  One 100 100.67 98.30 93.03 91.50 Comparative Example  2 100 100.50 99.58 98.65 99.51 Example  One 100 99.30 96.30 94.90 93.67 Example  2 100 98.25 99.77 99.51 96.95 Example  3 100 99.68 99.21 96.57 98.51

As a result, as shown in Table 5 above, the concentrations of Examples 1 to 3 and Comparative Examples 1 and 2 showed no cytotoxicity even at the highest concentration of 100 ug / ml, and thus it can be used as a safe raw material without toxicity Respectively.

Test Example 5: Antioxidant efficacy test: DPPH radical scavenging activity

DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging activity of the ginseng extract obtained from Example 1 and Comparative Example 1 was measured. Free radicals generally act as a pathologic factor that causes the skin to dry out by destroying the skin moisturizing layer and reacting to external stimuli, which is sensitive to external stimuli. The antioxidant activity of the extract of ginseng complex according to the present invention can be confirmed by free radical scavenging ability through DPPH experiment. First, the ginseng extract obtained in Example 1 and Comparative Example 1 was diluted to various concentrations, and samples were prepared. 100 μM DPPH (1,1-diphenyl-2-picrylhydrazyl) Min, and the amount of DPPH was measured at 520 nm in a 96-well plate. At this time, the case where no sample was added was used as a control group. The degree of antioxidation by the DPPH free radical scavenging activity (%) measurement method is shown in Equation 2 below.

&Quot; (2) &quot;

Free radical scavenging ability (%) = 100- (absorbance of the group treated with the sample / absorbance of the group not treated with the sample X 100)]

Free radical scavenging (%) density( ug / ml) 0 10 25 50 100 sample Comparative Example  One 0.77 22.43 34.43 56.00 77.47 0.78 20.54 29.51 30.64 30.82 0.77 30.28 59.58 70.51 75.90 Example  One 0.80 38.10 63.53 90.93 92.87 0.79 45.51 89.56 90.35 92.82

As a result, as shown in Table 6 and FIG. 3, when the ginseng extract was treated at different concentrations, the DPPH scavenging activity was significantly higher in Example 1 than in Comparative Example 1, indicating a high antioxidative activity.

<Test Example 6> Wrinkle-improving efficacy test: MMP-1 inhibitory activity

MMP-1 (Matrix Metallo peptidase-1) inhibitory activity of the ginseng extract obtained from Example 1 and Comparative Example 1 was measured. At this time, the amount of mRNA expression was examined by RT-PCR. Human fibroblasts were inoculated into each well of a 6-well plate to be 2 × 10 ⁵ cells and cultured in a 5% CO ₂ environment at 37 ° C. for 24 hours. When confluency of about 80% was reached, the cells were washed 3 times with phosphate buffered saline (PBS) and irradiated with ultraviolet light to induce skin aging. At this time, ultraviolet rays were irradiated with a total of 6.5 mJ / cm 2 using an ultraviolet A lamp (Sankyo Denki FL8BL lamp, Sankyo Denki Co., Japan). Immediately after UV irradiation, the cells were washed with phosphate buffer and the ginseng extract obtained in Example 1 and Comparative Example 1 was added to DMEM medium containing no fetal bovine serum at concentrations of 0, 10, 25, 50, 100, and 200 ug / ml Followed by further culture for 24 hours. RNA was isolated using the sample-treated cells, and the RNA was isolated using an RNA isolation kit (intron). CDNA was prepared by inserting MuLV reverse transcriptase and 0.5 μg of 1 mM dNTP into the obtained RNA. MMP-1 primer (SEQ ID NO: 1: F 5'-AAAGGGAATAAGTACTGGGC-3 'and SEQ ID NO: 2: R 5 '-ATTCCAGGAAAGTCATGTG-3') was amplified by real-time PCR (Real Time PCR).

MMP -1 inhibition (%) density( ug / ml) 0 10 25 50 100 sample Comparative Example  One 0.50 20.27 25.27 29.27 33.07 0.49 10.50 13.58 11.69 11.55 0.49 19.51 20.21 25.98 56.23 Example  One 0.50 32.57 44.63 52.33 72.00 0.51 40.68 62.39 80.54 82.35

As a result, when Example 1 and Comparative Example 1 were treated by concentration as shown in Table 7 and FIG. 4, the expression of MMP-1 increased by ultraviolet irradiation was remarkably decreased in Example 1 compared to Comparative Example 1 It was confirmed that there was an effect of improving skin wrinkles.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereto will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

<110> KANG, hee cheol <120> Method for producing ginseng extract with enhanced ginsenoside          Rg3, F1, C-K, Rh2 and PPT content <130> P16131 <160> 2 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MMP 1 primer F <400> 1 aaagggaata agtactgggc 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MMP 1 primer R <400> 2 aattccagga aagtcatgtg 20

Claims (10)

Enzymatic treatment of ginseng and subsequent drying;
Expanding dried ginseng; And
Wherein the ginsenoside Rg3, F1, CK, Rh21, and PPT are increased by adding the ginsenoside Rg3, F1, CK, Rh21 and PPT.
[Claim 2] The ginseng according to claim 1, wherein the ginseng is a mixture of leaves, fruits and roots at a mixing ratio of 0.5-1.0: 0.5-1.0: 0.5-5.0, A method for producing ginseng extract having enhanced contents.
[Claim 2] The method according to claim 1, wherein the enzyme is pectinase and cellulase, wherein the content of ginsenosides Rg3, F1, CK, Rh21 and PPT is increased.
The method according to claim 3, wherein the content of ginsenosides Rg3, F1, CK, Rh21 and PPT is increased by further adding lactose during the enzyme treatment.
The method according to claim 1, wherein the enzyme treatment comprises adding 0.1 to 1.0 part by weight of pectinase and 0.5 to 1.0 part by weight of a cellulase to 100 parts by weight of ginseng, Wherein the content of ginsenosides Rg3, F1, CK, Rh21 and PPT is increased.
2. The method according to claim 1, wherein the expanding step is performed at a pressure of 5.0 to 10.0 kgf / cm &lt; ² &gt;, wherein the content of ginsenosides Rg3, F1, CK, Rh21 and PPT is increased.
The method according to claim 1, wherein the extracting step comprises fractionating ginsenosides Rg3, F1, and Rg2 by extracting the expanded ginseng with ethanol (EtOH), and then extracting the extract using HP20 porous resin and ethanol (EtOH) Wherein the content of CK, Rh21 and PPT is increased.
A ginseng extract prepared by the method of any one of claims 1 to 7.
[Claim 9] The ginseng extract of claim 8, wherein the ginseng extract has skin antioxidant and wrinkle improving effect.
A cosmetic composition comprising the ginseng extract of claim 8.

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